Abstract

In vivo reprogramming of somatic cells into induced pluripotent stem cells (iPSC) holds vast potential for basic research and regenerative medicine. However, it remains hampered by a need for vectors to express reprogramming factors (Oct-3/4, Klf4, Sox2, c-Myc; OKSM) in selected organs. Here, we report OKSM delivery vectors based on pseudotyped Adeno-associated virus (AAV). Using the AAV-DJ capsid, we could robustly reprogram mouse embryonic fibroblasts with low vector doses. Swapping to AAV8 permitted to efficiently reprogram somatic cells in adult mice by intravenous vector delivery, evidenced by hepatic or extra-hepatic teratomas and iPSC in the blood. Notably, we accomplished full in vivo reprogramming without c-Myc. Most iPSC generated in vitro or in vivo showed transcriptionally silent, intronic or intergenic vector integration, likely reflecting the increased host genome accessibility during reprogramming. Our approach crucially advances in vivo reprogramming technology, and concurrently facilitates investigations into the mechanisms and consequences of AAV persistence.

Details

Title
AAV vector-mediated in vivo reprogramming into pluripotency
Author
Senís, Elena 1 ; Mosteiro, Lluc 2 ; Wilkening, Stefan 3 ; Wiedtke, Ellen 4 ; Nowrouzi, Ali 3 ; Afzal, Saira 3 ; Fronza, Raffaele 5 ; Landerer, Henrik 6 ; Abad, Maria 7 ; Niopek, Dominik 8   VIAFID ORCID Logo  ; Schmidt, Manfred 5 ; Serrano, Manuel 9 ; Grimm, Dirk 10 

1  Virus-Host Interaction Group, Department of Infectious Diseases/Virology, Heidelberg University Hospital, Cluster of Excellence CellNetworks, Heidelberg, Germany; BioQuant, University of Heidelberg, Heidelberg, Germany; Cellular Plasticity and Cancer Group, Vall d’Hebron Institute of Oncology (VHIO), Barcelona, Spain 
2  Tumor Suppression Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain 
3  Department of Translational Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ), Heidelberg, Germany 
4  Virus-Host Interaction Group, Department of Infectious Diseases/Virology, Heidelberg University Hospital, Cluster of Excellence CellNetworks, Heidelberg, Germany; BioQuant, University of Heidelberg, Heidelberg, Germany 
5  Department of Translational Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ), Heidelberg, Germany; GeneWerk GmbH, Heidelberg, Germany 
6  Virus-Host Interaction Group, Department of Infectious Diseases/Virology, Heidelberg University Hospital, Cluster of Excellence CellNetworks, Heidelberg, Germany; BioQuant, University of Heidelberg, Heidelberg, Germany; Institute for Pharmacy and Molecular Biotechnology (IPMB), Bioinformatics Division, University of Heidelberg, Heidelberg, Germany 
7  Cellular Plasticity and Cancer Group, Vall d’Hebron Institute of Oncology (VHIO), Barcelona, Spain 
8  BioQuant, University of Heidelberg, Heidelberg, Germany; Institute for Pharmacy and Molecular Biotechnology (IPMB), Bioinformatics Division, University of Heidelberg, Heidelberg, Germany; Theoretical Bioinformatics Division, German Cancer Research Center, Heidelberg, Germany 
9  Tumor Suppression Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain; Institute for Research in Biomedicine (IRB Barcelona), Barcelona Institute of Science and Technology (BIST), Catalan Institution for Research and Advanced Studies (ICREA), Barcelona, Spain 
10  Virus-Host Interaction Group, Department of Infectious Diseases/Virology, Heidelberg University Hospital, Cluster of Excellence CellNetworks, Heidelberg, Germany; BioQuant, University of Heidelberg, Heidelberg, Germany; German Center for Infection Research (DZIF), Partner site Heidelberg, Heidelberg, Germany 
Pages
1-14
Publication year
2018
Publication date
Jul 2018
Publisher
Nature Publishing Group
e-ISSN
20411723
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41467-018-05059-x
ProQuest document ID
2067108599
Copyright
© 2018. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.